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Issue 5, 2015
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Specific in situ hepatitis B viral double mutation (HBVDM) detection in urine with 60 copies ml−1 analytical sensitivity in a background of 250-fold wild type without DNA isolation and amplification

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Abstract

We have examined in situ detection of hepatitis B virus 1762T/1764A double mutation (HBVDM) in urine using a (Pb(Mg1/3Nb2/3)O3)0.65(PbTiO3)0.35 (PMN-PT) piezoelectric plate sensor (PEPS) coated with a 16-nucleotide (nt) probe DNA (pDNA) complementary to the HBVDM. The in situ mutation (MT) detection was carried out in a flow with the PEPS vertically situated at the center of the flow in a background of wild type (WT). For validation, this detection was followed by detection in the mixture of MT fluorescent reporter microspheres (FRMs) (MT FRMs) and WT FRMs that emitted different fluorescence colours and were designed to specifically bind to MT and WT, respectively. At 30 °C and 4 ml min−1, a PEPS was shown to specifically detect HBVDM in situ with 60 copies ml−1 analytical sensitivity in a background of clinically-relevant 250-fold more WT in 30 min without DNA isolation, amplification, or labelling as validated by the visualization of the captured MT FRMs and WT FRMs following FRM detection where the captured MT FRMs outnumbered the WT FRMs by a factor of 5 to 1.

Graphical abstract: Specific in situ hepatitis B viral double mutation (HBVDM) detection in urine with 60 copies ml−1 analytical sensitivity in a background of 250-fold wild type without DNA isolation and amplification

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Publication details

The article was received on 17 Oct 2014, accepted on 05 Jan 2015 and first published on 05 Jan 2015


Article type: Paper
DOI: 10.1039/C4AN01885K
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Citation: Analyst, 2015,140, 1590-1598
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    Specific in situ hepatitis B viral double mutation (HBVDM) detection in urine with 60 copies ml−1 analytical sensitivity in a background of 250-fold wild type without DNA isolation and amplification

    C. E. Kirimli, W. Shih and W. Y. Shih, Analyst, 2015, 140, 1590
    DOI: 10.1039/C4AN01885K

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